Objective To develop an erythrocyte-based delivery system for butyrylcholinesterase(BChE)that is capable of prophylaxis against organophosphorus nerve agents.Methods Recombinant BChE was produced and analyzed for oligomerization via polyacrylamide gel electrophoresis(PAGE)and Western blotting.A modified hypotonic preswelling method was employed to prepare BChE-loaded erythrocytes.The drug loading capacity and encapsulation efficiency were quantified using enzyme-linked immunosorbent assay(ELISA).Catalytic activity was assessed in vitro with an activity detection kit.The system was characterized via scanning electron microscopy(SEM),flow cytometry and a hematology analyzer.Results Recombinant BChE predominantly existed as dimers(85％dimer,15％monomer).The optimized volume ratio of erythrocytes to hypotonic solution was determined as 1:7.Compared with native and empty erythrocytes,BChE-loaded erythrocytes exhibited significantly higher catalytic activity(P＜0.001).The mean corpuscular volume of BChE-loaded erythrocytes increased(P＜0.001),while the mean content of corpuscular hemoglobin and hemoglobin in erythrocytes per 100 mL decreased(P＜0.001).SEM revealed no morphological differences(biconcave disc shape).Hypotonic preswelling moderately increased erythrocyte apoptosis(P＜0.001),but no statistical difference was observed between BChE-loaded and hypotonic-treated erythrocytes(P＞0.05).CD47 expression remained unchanged compared to native erythrocytes(P＞0.05).Conclusion The modified hypotonic preswelling method can generate BChE-loaded erythrocytes that retain the characteristics of native erythrocytes while conferring catalytic activity,offering a novel strategy for clinical intervention against organophosphorus poisoning.

Objective To explore the safety and feasibility of the application of day surgery management model under enhanced recovery after surgery(ERAS)concept in patients undergoing gynecologic laparoscopic surgery.Methods A non-randomized concurrent control trial was conducted on the patients who underwent laparoscopic surgery in our department from January to August 2021.A total of 92 patients admitted on odd date were assigned into Ward B of our department and served as the control group,and another 96 patients hospitalized on even date were subjected into Ward A and served as the observation group.The control group was given the routine treatment schedule,including the relevant examinations after admission and general operation procedure during hospitalization.The observation group was under a day surgery management model based on the concept of ERAS,with aid of a day surgery team and optimized perioperative management measures,including pre-hospital rehabilitation,shortening water fasting before surgery,multi-mode analgesia,preventive antiemesis,intraoperative warmth,prevention of deep vein thrombosis,immediate postoperative eating and activity,and follow-up after discharge.Postoperative subjective comfort,intestinal function recovery,social and economic benefits,postoperative complications and inflammatory indicators were compared between the 2 groups.Results In 0～6,7～12 and 13～24 h after operation,the scores of thirst,hunger,nausea,pain,abdominal distension and pharyngeal discomfort were significantly lower in the observation group than the control group(P＜0.01).The observation group had obviously shorter length of hospital stay and ealier bowel sound recovery and first anal exhaust than the control group(P＜0.01).No postoperative complication,such as fall,unplanned secondary operation or wound infection was observed in both groups.The postoperative inflammatory indicators,including procalcitonin(PCT),neutrophil percentage(Neu％)and white blood cell count(WBC)were all in the normal ranges in the 2 groups at 24 h and 3 and 7 d after surgery.Statistical differences were found in firstly postoperative mobilization,length of hospital stay,hospitalization cost and patient satisfaction between the 2 groups(P＜0.01).Conclusion ERAS-based day surgery management model has the advantages of shortening hospital stay,reducing medical costs,promoting postoperative rehabilitation,and improving the comfort and satisfaction in patients undergoing gynecologic laparoscopic surgery.

Objective:To analyze the electroencephalogram (EEG) features of patients with Alzheimer’s disease (AD) and mild cognitive impairment (MCI), and to combine the characteristics for classification and prediction.Methods:One hundred and thirty-five patients attending the Department of Neurology at the General Hospital of Tianjin Medical University were enrolled, including 34 patients with AD, 67 patients with MCI, and 34 healthy control (HC). The electroencephalogram signals of these patients in the resting state were collected and preprocessed. Relative power spectral density features and sample entropy features on a multi-band scale were extracted to compare the whole-brain differences in electroencephalogram features among the 3 groups of subjects, and then subdivided into brain regions and individual leads for in-depth analysis. The above two features were fused to classify and predict AD, MCI, and HC by support vector machine (SVM).Results:The frontal regions had higher δ relative power spectral densities than the other regions, and the occipital and temporal regions showed relatively lower distributions. θ-Band relative power spectral densities had a more even distribution of sizes across brain regions. α-Band relative power spectral densities were concentrated in the occipital lobe, while β-band relative power spectral densities were mainly concentrated in the parietal and temporal lobes. Except for the central lobe, the δ-band relative power spectral densities of the AD group were higher than those of the MCI group ( P < 0.05) and HC group ( P < 0.01) in all brain regions and the whole brain. θ-band relative power spectral densities of the AD group were higher than those of the MCI gourp ( P < 0.001) and HC group ( P < 0.001) in the whole brain and in all brain regions. α-Band relative power spectral densities of the AD group were lower than those of the other groups only in the temporal lobe (all P < 0.05). The relative power spectral density of the β-band in the AD group was higher than that of the other groups in the whole brain and in all brain regions ( P < 0.05, 0.01, 0.001). The difference in the relative power spectral density of the δ-band in the C3 lead in the central lobe of the AD and HC groups was statistically significant ( P < 0.05). The relative power spectral density of the γ-band in the temporal lobe was higher than that in the other regions of the AD group, the MCI group, and the HC group. The relative power spectral density of the γ-band in the T3 lead in the AD group was significantly lower than that in the T4 lead. The average entropy of samples in the whole brain and in each brain region was lower than that in the HC group in the AD and MCI groups (all P < 0.05). The entropy of the samples at lead C3 in the AD group was lower than that in the MCI group ( P < 0.05). The differences between the relative power spectral density, sample entropy, and the actual data classification evaluation indexes (accuracy rate, precision rate, recall rate, and F1 score) that fused the two features, and the rearranged data were all statistically significant (all P < 0.001). When the relative power spectral density feature and the sample entropy feature were fused in the classification features, the best classification prediction was achieved, with an accuracy rate of 80%, a precision rate of 78%, a recall rate of 78%, and the F1 score of 79%. Conclusions:Relative power spectral density and sample entropy analysis can reveal the abnormalities of electroencephalogram activities of AD and MCI patients from different perspectives (linear and nonlinear), and the combination of these two features in classification prediction can improve the classification effect.

Objective To investigate the pathological damage to and inflammation of different intestinal segments in a rat model of severe hemorrhage,and to explore the effect of polarization of intestinal macrophage on the pathophysiology of intestinal inflammation.Methods Male Wistar rats were randomly divided into two groups:the sham operation group and hemorrhage group.In the hemorrhage group,40％of the total blood volume was lost in 25-30 minutes,while in the sham operation group,only the femoral artery and vein were intubated without bleeding.The rats were killed at 0,3,6,12 and 24 hours.The entire intestine was isolated quickly,and sections of the intestine were cut at the duodenum,jejunum,ileocecal junction,colon and rectum for histopathological evaluation.ELISA was adopted to determine related inflammation factors while multi-color immunohistochemistry was used to calculate macrophage surface markers.The data was statistically analyzed.Results(1)Compared with the sham group,there was no significant difference in colon histology at 3 h and 6 h,but significant difference was detected in rectum scores only at 24 h.The scores of other intestinal segments were significantly different at each time point.The severity of ileocecal and colonic lesions after bleeding increased with time.The duodenum,jejunum and ileocecum were more critically injured at 3 h than the rectum at 6 h.The injury to the duodenum,jejunum,ileum and colon was much more pronounced than to the rectum at 12 h.(2)The expressions of TNF-α and IL-1β in the rectum were increased significantly at 12 h post operation.The expressions of IL-1β,TNF-α in the jejunum increased obviously at 3 h and 6 h,respectively.(3)Three hours after severe bleeding,the level of macrophages in the jejunum and ileocececal area increased significantly,and the percentage of M1 macrophages was higher.After 6 hours,the proportion of M2 macrophages in the jejunum and M1 macrophages decreased significantly.After 3 hours,the percentage of M1 macrophages in the colon decreased,but that of M2 macrophages increased.The proportion of M2 polarized macrophages in the duodenum and rectum increased at 3 h after severe bleeding but decreased at 6 h.Conclusion Pathological damage to intestinal sections after bleeding varies depending on the time,and is correlated with the inflammatory level of macrophages.

Objective To explore the effects of prucalopride(PRUC)on the intestinal function during the perioperative period of robot-assisted laparoscopic radical cystectomy(RARC)and urinary diversion.Methods A total of 75 patients undertaking RARC with urinary diversion(orthotopic neobladder or ileal bladder)in Nanjing Drum Hospital during Jan.and Dec.2021 were divided into PRUC group(n=28)and control group(n=47)according to whether they took PRUC or not.Postoperative intestinal ventilation time and defecation time,drainage tube retention time,tolerance time for first intake of semi-flow food,postoperative hospital stay,and incidence of complications were observed and recorded in the two groups.Postoperative C-reactive protein(CRP)and neutrophil/lymphocyte ratio(NLR)were compared.Results The PRUC group had shorter intestinal ventilation time and defecation time[(47.14±16.31)h vs.(74.04±35.33)h,P＜0.01;(86.14±30.47)h vs.(123.57±79.12)h,P=0.02],smaller change of ΔCRP and ΔNLR[(79.99±29.71)mg/L vs.(127.75±56.98)mg/L;(9.24±6.43)vs.(16.11±9.90),P＜0.01].All complications were minor,the incidence of intestinal obstruction in PRUC group tended to decrease within 90 days after operation(P=0.38),and there was no significant difference in other complications between the two groups(P＞0.05).Conclusion The perioperative use of PRUC in RARC with urinary diversion is safe and effective,which can promote the recovery of intestinal function after operation.

Objective Based on the gene expression omnibus(GEO)database,bioinformatics methods were employed to analyze the expression characteristics of hypoxia-related differentially expressed genes(HRDEGs)in ischemic stroke,and key genes were screened,to provide important support for a deeper understanding of ischemic stroke.Methods The GSE16561 and GSE58294 datasets were downloaded from the GEO database,and Python software was used for data integration.The Combat method was employed to eliminate batch effects while retaining disease grouping characteristics.Principal component analysis was conducted to reduce dimensionality of the data before and after batch effect removal,and intraclass correlation coefficient(ICC)testing was performed on the ischemic stroke and normal control groups.Gene set enrichment analysis(GSEA)and single-sample GSEA were conducted on the merged and batch effects eliminated dataset,with a nominal P-value(NOM P-val)＜0.05 and false discovery rate P-value(FDR P-val)＜0.25 used as criteria to select significantly different gene sets.Differential expression genes between the ischemic stroke samples and normal control samples after merging and eliminating batch effects of the GSE16561 and GSE58294 datasets were identified using R software,with an absolute value of log2 gene expression fold change(FC)≥0.58 and adjusted P-value(Padj)＜0.05 as selection criteria.Intersection with hypoxia-related genes obtained from the National Center for Biotechnology Information(NCBI)in the United States yielded the HRDEGs.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analyses were performed on the HRDEGs,and the STRING database was used to construct a protein-protein interaction network of differentially expressed genes.The top 10 key genes were filtered using Cytoscape 3.8 software.Results The ICC analysis results showed excellent consistency in the ischemic stroke and normal control samples after batch effect removal,with ICC values of 0.94 and 0.98 for the GSE16561 and GSE58294datasets,respectively.GSEA results demonstrated significant enrichment of 34 gene sets in the stroke samples in the newly merged and batch effects removed dataset from GSE16561 and GSE58294,leading to the identification of 404 differentially expressed genes(all with Padj＜0.05),including 354 upregulated genes and 50 downregulated genes.Intersection with hypoxia-related genes yielded 64 HRDEGs.GO enrichment analysis indicated significant enrichment of HRDEGs in vesicle lumen,cytoplasmic vesicle lumen,secretory granule lumen,with molecular functions such as amide binding,peptide binding,phospholipid binding,and enzyme inhibitor activity.These genes are primarily involved in the positive regulation of cytokine production,regulation of immune response,response to bacterium-derived molecules,and response to lipopolysaccharide,among other biological processes.KEGG enrichment analysis revealed enrichment of HRDEGs in pathways related to lipid and atherosclerosis,Salmonella infection,neutrophil extracellular trap formation,nucleotide-binding oligomerization domain-like receptor signaling pathway,protein glycosylation in cancer,tuberculosis,and necroptosis.Based on the protein-protein interaction network,10 key genes were identified,including arginase1(ARG1),caspase1(CASP1),interleukin1 receptor type 1(IL-1R1),integrin subunit alpha M(ITGAM),matrix metalloproteinase9(MMP9),prostaglandin-endoperoxide synthase 2(PTGS2),signal transducer and activator of transcription 3(STAT3),Toll-like receptor2(TLR2),TLR4,and TLR8.Conclusion This study has identified 10 key genes associated with ischemic stroke and hypoxia through bioinformatics mining,which maybe provid potential targets for subsequent research and diagnostic and therapeutic interventions.

Objective: To explore the differential expression profiles of DNA methylation sites/regions and potential molecular mechanisms in the peripheral blood of coronary heart disease (CHD)-induced unstable angina pectoris patients with or without Qi deficiency and blood stasis syndrome, and to provide scientific evidence for the conbination of disease and syndrome. Methods: According to the pre-determined inclusion and exclusion criteria, the study subjects were enrolled and divided into two groups namely CHD-induced unstable angina group (G group) and healthy control group (J group) to conduct “disease” analysis, while G group was further divided into Qi deficiency and blood stasis syndrome group (case group) and non-Qi deficiency blood stasis syndrome group (control group) to perform “syndrome” analysis. The general data and clinical information of the study subjects were collected. The peripheral venous blood was extracted on an empty stomach, and the Illumina Infinium MethylationEPIC BeadChip (850K methylation chip) was used to detect the differential expressionprofiles of DNA methylation in each group, ChAMP software (V 2.14.0) was used for the differential methylation data analysis, with a threshold of the adjusted P value (adj.P.val) < 0.01. Gene Ontology (GO) and Kyoto Encyclopedia of Genomes (KEGG) were employed for the functional and pathway enrichment analyses of related mapped genes. Results: A total of 263 differentially methylated CpG positions (DMPs) were screened out between G and J groups, including 191 hypermethylated positions such as cg05845204 and cg08906898, and 72 hypomethylated positions such as cg26919182 and cg13149459. These positions were mainly mapped to 148 genes encompassing RNA binding motif protein 39 (RBM39), acetyl-CoA acyltransferase 2 (ACAA2), protein phosphatase 1 regulatory subunit 12B (PPP1R12B), and the dual-specificity tyrosine phosphorylation-regulated kinase 2 (DYRK2). GO functional enrichment analysis revealed that the genes of the DMPs were primarily enriched in protein localization to chromosomes, regulation of cell morphogenesis, negative regulation of calcium-mediated signals, etc. KEGG pathway analysis suggested that the genes were mainly enriched in fatty acid metabolism and endocytosis pathways. In addition, a total of 23 differential methylation regions (DMRs) were identified, with overlapping genes such as transmembrane protein 232 (TMEM232), ribosomal protein large P1 (RPLP1), peroxisomal biogenesis factor 10 (PEX10), and forkhead box N3 (FOXN3) recognized. It was found that GO functions were mainly enriched in the negative regulation of Ras protein signal transduction, small GTPase-mediated signal transduction, negative regulation, etc. A total of 1 703 differential methylation sites were screened out between case and control groups, including 444 increased methylation positions such as cg05573767 and 1 259 decreased methylationpositions such as cg19938535, and cg03893872. These positions were mapped to 1 108 genes such as ribosomal protein S6 kinase A2 (RPS6KA2), leucine rich repeat containing 16A (LRRC16A), and hedgehog acyltransferase (HHAT). According to the GO functional enrichment analysis, the genes relating to the DMPs were mainly enriched in biological functions such as transmembrane receptor protein serine/threonine kinase signaling pathway and axonogenesis. The KEGG pathway enrichment analysis suggested the involvement of Rap1 signaling pathway, adenosine 5’-monophosphate-activated protein kinase (AMPK) signaling pathway, etc. A total of 21 DMRs were identified, including 22 overlapping genes such as mucin 4 (MUC4), three prime repair exonuclease 1 (TREX1), and LIM homeobox 6 (LHX6). GO analysis demonstrated that the genes primarily participated in molecular functions such as positive regulation of transmembrane transport, regulation of fatty acid metabolism, and copper ion binding. Conclusion This study reveals the methylation patterns of DMPs and DMRs in patients with Qi deficiency and blood stasis syndrome caused by CHD-induced unstable angina pectoris. Potential epigenetic regulation of fatty acid metabolism, Rap1 signaling, and other molecular functions are involved in the development of CHD between the "disease" and "syndrome".

Objective To investigate the change of infusion particles and PH value of rifampicin for injection under dif-ferent configuration and application conditions,and to provide the best configuration method and the best infusion quality control strategy for rifampicin for injection.Methods The particle number and PH value of rifampin for injection were investigated and measured under different configuration and application conditions,and the factors affecting the formation of particles were ana-lyzed.Results The stable pH value of Rifampicin for injection was 7.2-8.1,and the number of particles was the least in the light filtering and 5%glucose 500 mL,and the number of particles was the least in 1～4 h after the configuration.Conclusion The optimal infusion quality control strategy is to filter rifampicin for injection by blocking light,500 mL of solute 5%glucose,1～4 h after configuration,and use of disposable precision filter infusion set.

Objective This study aimed to conduct a systematic review and meta-analysis of randomized controlled trials(RCTs)assessing the clinical efficacy of acupuncture for chronic stable angina(CSA).Methods PubMed,The Cochrane Library,Embase,CNKI,VIP,WanFang Data,CBM databases,ClinicalTrial.gov and Chinese Clinical Trial Registry were electronically searched to collect randomized controlled trials(RCTs)of acupuncture for CSA and only RCTs that included acupuncture as the sole or combination of acupuncture in the treatment of CSA.The retrieval period was from the establishment of the database to May 19,2022.The primary outcome measure was the frequency of angina attacks,and the secondary outcome measures were angina efficacy,total effective rate of ECG improvement,nitroglycerin use after treatment,patient-perceived overall effectiveness,performance on the Six-Minute Walk Test(6-MWT),depression as measured by the anxiety as measured by the Self-Rating Anxiety Scale(SAS),Self-Rating Depression Scale(SDS),adverse effects.Results Twelve studies with a total of 1605 case subjects were included and grade quality of evidence reviews had 1 outcomes as medium quality evidence,5 outcomes as low quality evidence and 4 outcomes as very low quality evidence.Acupuncture treatment resulted in significantly greater reductions in angina attacks,visual analog scale(VAS),6-minute walk test,angina efficacy,electrocardiogram improvement rate,angina efficacy,total clinical response rate.Conclusion The efficacy of acupuncture combined with western medicine in treating CSA is better than that of Western medicine alone,thanks to restricted by the number and quality of included researchs,higher quality studies are required to prove above conclusions.